Abstract
Purpose :
Investigate the effect of pore size and shape of 3D biomimetic scaffolds on the proliferation and fibronectin expression of human trabecular meshwork (hTM) cells.
Methods :
Scaffolds were constructed using collagen and the glycosaminoglycans chondroitin sulfate (CS) and/or hyaluronic acid (HA), components found in the native TM extracellular matrix (ECM). Pore alignment molds were used to produce either aligned or random pores. Different freezing temperatures (-80C and -196C) were used to produce large or small pores, respectively. hTM cells were seeded onto the surface of these scaffolds and cultured for up to 4 weeks. Proliferation was measured using a metabolic assay. Expression of fibronectin was measured using qPCR and visualized by confocal microscopy.
Results :
Fabricated scaffolds display aligned or random pores, with average pore sizes of either 161 ± 70 µm (large) or 10 ± 5 µm (small) as measured by SEM images. hTM cells proliferated on these scaffolds for up to 4 weeks in culture. Cells grown on the large pore scaffolds proliferated more, regardless of the GAG content, and exhibited greater fibronectin expression and distribution. Additionally, it was found that fibronectin expression was higher for hTM cells cultured on scaffolds that contained either CS or HA compared with collagen only. Small pore scaffolds show less hTM cells migration into the matrix and their fibronectin deposition was limited to the top layers of the scaffold.
Conclusions :
Our results confirm that hTM cells grow and proliferate on collagen-GAG scaffolds. The underlying scaffold architecture affects the growth, gene expression, and protein deposition of these hTM cells. Large pores allow for hTM cells to grow and secrete increased amounts of their own ECM. The presence of GAG, either CS or HA, in the scaffolds increased the expression of fibronectin.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.